Search

X-ray phase-contrast computed tomography imaging based on the Talbot grating interferometer is described, and the way it can visualize the polyglycolic acid scaffold, including its microfibres, after implantation into the injured spinal cord is shown., Tissue engineering strategies for spinal cord repair are a primary focus of translational medicine after spinal cord injury (SCI). Many tissue engineering strategies employ three-dimensional scaffolds, which are made of biodegradable materials and have microstructure incorporated with viable cells and bioactive molecules to promote new tissue generation and functional recovery after SCI. It is therefore important to develop an imaging system that visualizes both the microstructure of three-dimensional scaffolds and their degradation process after SCI. Here, X-ray phase-contrast computed tomography imaging based on the Talbot grating interferometer is described and it is shown how it can visualize the polyglycolic acid scaffold, including its microfibres, after implantation into the injured spinal cord. Furthermore, X-ray phase-contrast computed tomography images revealed that degradation occurred from the end to the centre of the braided scaffold in the 28 days after implantation into the injured spinal cord. The present report provides the first demonstration of an imaging technique that visualizes both the microstructure and degradation of biodegradable scaffolds in SCI research. X-ray phase-contrast imaging based on the Talbot grating interferometer is a versatile technique that can be used for a broad range of preclinical applications in tissue engineering strategies.

The goal of regenerative medicine is to achieve histological and functional recovery to the level of the original tissue. For this purpose, we have developed a biodegradable scaffold to create cell-free in-situ tissue-engineered vasculature (iTEV) with good long-term results. However, the regeneration process of vascular smooth muscle cells (VSMCs) over time has yet to be examined. To evaluate the regeneration ability of VSMCs, the inferior vena cava of experimental animals was replaced with iTEV, and tested at 1, 3, 6, 12, and 24 months (n = 6 each) after implantation. Six animals were enrolled to compare 24-month iTEV and native vasculature in single individual samples. There were no complications throughout the study. Immunohistology, protein expression analysis, and biochemical findings indicate that iTEV can gradually regenerate and develop into a mature vessel within 24 months using our biodegradable scaffold. These results provide a time course for the regeneration of VSMCs within the tissue-engineered vascular autograft constructed using a biodegradable scaffold.

Poly(L-lactic acid) felt (PLLA felt) was prepared for reconstruction of the rotator cuff in animal models. Small changes were found in the tensile strength of both the cultured PLLA felt and the PLLA felt implanted on the paravertebral muscle of rabbits up to 16 postoperative weeks. The stiffness of the felt implanted on the muscle from 6 to 16 weeks showed a statistically significant increase. When the infraspinatus tendons of beagle dog were reconstructed with the PLLA felt, the ultimate strength of PLLA felt increased threefold, and the stiffness increased fivefold by 16 postoperative weeks compared to that of the initial PLLA felt. They were statistically significant (p < 0.01). All the implanted specimens ruptured at the junction between the bone and the PLLA felt. Histological examination demonstrated infiltration of fibrous tissue into the interstices of the PLLA felt fibers. Connection between the infraspinatus tendon and the PLLA felt was tight with the formed scar tissue, but the connective tissue between the bone and PLLA felt fibers was sparse even at 16 and 32 postoperative weeks. A few deteriorated PLLA felt fibers were observed at 32 postoperative weeks. It was concluded that the degradation rate of PLLA felt was low and the tensile recovery of the PLLA felt graft in beagle dogs was excellent. Thus, PLLA felt might be a useful bioabsorbable material for rotator-cuff reconstruction.

Local drug delivery is an attractive approach to the associated problems of percutaneous transluminal coronary angioplasty (PTCA), including arterial injury. The objective of the present research was to deliver a high concentration of a potent anti-thrombin agent, argatroban (ARG), to the vessel wall in order to reduce arterial injury. Local delivery was accomplished by the ionic attachment of drug particles to a modified balloon surface. Surface graft polymerization of ionic monomers to a high-density poly(ethylene) (PE) substrate was performed utilizing ultra-violet (UV) methods. Acrylic acid (AAc) and 2(dimethylamino) ethyl methacrylate (DMAEMA) were successfully grafted onto PE surfaces. Surface grafting was verified by contact angle, X-ray photoelectron spectroscopy, and zeta potential measurements. The amount of ARG adsorbed onto the modified PE surface was highly dependent on the pH of the drug media for both anionic and cationic grafted monomers. The efficacy of local drug delivery to the arterial wall was analyzed using drug-immobilized PE balloon catheters in the rabbit common carotid artery model. High concentrations of ARG (280 nmol/g tissue) were found within the ballooned arterial segment immediately after angioplasty, followed by a decrease after blood flow was restored.

Although fibrin glue has been widely used as a surgical adhesive, its components, fibrinogen and thrombin, obtained from human blood are not completely free from the risk of virus infection due to acquired immune deficiency and hepatitis. Recently, we have reported that a polymer pair composed of gelatin and poly(L-glutamic acid) (PLGA) promptly forms a gel and can firmly bond to soft tissues when crosslinked with the aid of water-soluble carbodiimide (WSC). The present study was undertaken to design a new PLGA-gelatin glue without using WSC. Two kinds of PLGA with molecular weights of 71 and 22 kDa were employed to prepare N-hydroxysuccinimide (NHS) activated derivatives. The NHS-activated PLGA could be synthesized at high yields and was found to be stable for an extended time without losing the ability to crosslink with gelatin when stored under a dry-cold condition. This NHS-activated PLGA could spontaneously form a gel with gelatin in an aqueous solution within a short time, comparable to a commercial fibrin glue, when gelation was allowed to proceed at pH 8.3. The NHS-activated PLGA prepared from PLGA with the molecular weight of 22 kDa could be readily dissolved at high concentrations and its ability to form a gel was maintained for more than 10 min when an acidic 8% NHS-activated PLGA solution was used. The bonding strength of PLGA gelatin glues with natural tissue was higher than that of fibrin glue. These findings strongly suggest that this combination of gelatin and NHS-PLGA is very promising as a surgical adhesive and may possibly replace fibrin glues prepared from human blood components.

Reconstructions of the intrathoracic trachea in 24 dogs were done with the use of 50 mm long collagen-conjugated tracheal prostheses. Omental wrapping was also done in 14 of the dogs (omentopexy group) to evaluate the efficacy of this option in comparison with results in the other 10 dogs (control group). All 24 dogs had uneventful postoperative courses and were killed at 4 weeks or 3, 6, or 12 months after the operation. Better epithelialization and fewer complications, such as mesh exposure and luminal stenosis, were observed in the omentopexy group than in the control group. Angiography and analysis of regenerated blood vessels revealed that vessel ingrowth had started within 4 weeks and that vessel formation reached its maximal point within 6 to 12 months in the omentopexy group. In contrast, revascularization of the subepithelial region in the control group was poor even after 3 months, and vessel formation continued for as long as 12 months. The differences between the two groups were considered to be mainly a result of the speed of blood vessel ingrowth into the regenerated mucosa. We conclude that our prosthesis can be used safely for intrathoracic tracheal reconstruction and that omental wrapping is a useful supplementary method that reduces the occurrence of complications. (J Thorac Cardiovac Surg 1997;113:691-700)

Immunoisolation, that is, enclosure of cells within a semipermeable membrane to protect them from immunological rejection, may enable the transplantation of cells without use of immunosuppressive drugs. Therefore, in addition to naturally-occurring ionic polymers, several synthetic nonionic polymers which can form dense and strong membranes in water have been studied as materials for immunoisolation. However, such nonionic polymers are required to be soluble in organic solvents which are mostly cytotoxic. In this report we describe enclosure of insulin-releasing cells into water-insoluble poly(2-hydroxyethyl methacrylate) and poly(2-hydroxyethyl methacrylate-co-methyl methacrylate) membranes using X-ray contrast medium as a solvent without use of any special apparatus. The contrast medium employed in our study is iopamidol aqueous solution. Insulin release was observed for 1 months when insulin-releasing cells were encapsulated into these membranes. The permeability of five solutes through the membranes prepared from the iopamidol aqueous solution was also studied to determine their potential immunoisolative efficacy.

Interventional radiology is becoming one of the standard treatments of arteriovenous malformation (AVM). Cyanoacrylate derivatives and polymer solutions are widely used to occlude the AVM nidus by their injection through a catheter, but they are far from satisfactory embolic liquids. For instance, cyanoacrylate derivatives sometimes glue the catheter to the artery, resulting in serious complications; in addition, the organic solvents used to dissolve polymers cause damage to the surrounding brain tissue of the AVM. Therefore, we attempted to develop embolic liquids by dissolving poly(2-hydroxyethyl methacrylate-co-methyl methacrylate) in Iopamiron with an addition of a small amount of ethyl alcohol. This new embolic liquid is not cytotoxic and is easily injected into the AVM through a thin, long catheter to effectively occlude the AVM.

We previously developed a cell-free, biodegradable scaffold for in-situ tissue-engineering vasculature (iTEV) in a canine inferior vena cava (IVC) model. In this study, we investigated application of this scaffold for iTEV of the pulmonary artery (iTEV-PA) in a canine model. In vivo experiments were conducted to determine scaffold characteristics and long-term efficacy. Biodegradable scaffolds comprised polyglycolide knitted fibers and an l-lactide and e-caprolactone copolymer sponge, with an outer glycolide and e-caprolactone copolymer monofilament reinforcement. Tubular scaffolds (8 mm diameter) were implanted into the left pulmonary artery of experimental animals (n = 7) and evaluated up to 12 months postoperatively. Angiography of iTEV-PA after 12 months showed a well-formed vasculature without marked stenosis, aneurysmal change or thrombosis of iTEV-PA. Histological analysis revealed a vessel-like vasculature without calcification. However, vascular smooth muscle cells were not well-developed 12 months post-implantation. Biochemical analyses showed no significant difference in hydroxyproline and elastin content compared with native PA. Our long-term results of cell-free tissue-engineering of PAs have revealed the acceptable qualities and characteristics of iTEV-PAs. The strategy of using this cell-free biodegradable scaffold to create relatively small PAs could be applicable in pediatric cardiovascular surgery requiring materials.

The manual seeding of cells onto a biodegradable scaffold by pipetting is an effective method of cell seeding. However, the widespread use and ultimate clinical utility of this technique is limited by operator variability. This study was conducted to evaluate an operator-independent vacuum-seeding method for use in an upcoming clinical trial. Using bone marrow-derived mononuclear cells, we achieved seeding comparable to manually seeded scaffolds in terms of cellular attachment, distribution, and viability in vacuum-seeded grafts at vacuum pressures of −25 to −50 mmHg. In conclusion, we describe an operator-independent seeding method for use in the clinical setting.

Limited information exists regarding the usefulness of bioabsorbable materials in the design of tracheal grafts. The aim of this study was to evaluate the feasibility of three bioabsorbable materials for use as artificial trachea. Three sets of grafts were prepared: Group 1 (n=6), knitted polyglactin 910 mesh; Group 2 (n=3), copolymer of L-lactide and epsilon-caprolactone sponge reinforced with polyglycoride fibers; and Group 3 (n=8), copolymer of L-lactide and epsilon-caprolactone sponge covered with knitted poly-L-lactide mesh. All grafts were internally reinforced with a titanium stent. A 10-cartilage-ring-length of canine mediastinal trachea was resected and replaced by a bioabsorbable prosthesis with the aid of an omental flap. In Groups 1 and 2, the patency rates decreased below 50% within two months after surgery. In Group 3, six of eight dogs maintained patency rates above 50% from 10 months to 2 years after surgery. Grafts prepared with a copolymer of L-lactide and epsilon-caprolactone sponge covered with knitted poly-l-lactide mesh (Group 3) can function for up to two years after surgery. These results provide evidence toward the feasibility of utilizing bioabsorbable materials as a tracheal prosthesis.

The authors evaluated the embolization effects on experimental aneurysm models with a liquid material. This material is a mixture of poly (HEMA-co-MMA), Iopamidol, water and a small amount of ethyl alcohol. We have satisfactorily treated over 30 cases of AVM with this material. Out of four solutions of different viscosity, we used the solution with the highest viscosity to prevent distal migration of the mixture. Glass models of aneurysms of 5 mm in diameter in pulsatile saline flow were embolized with the mixture under various conditions. When an IDC was placed in the aneurysm models, the mixture obliterated them completely and did not migrate out from the aneurysm models. In this experimental study, it is suggested that the liquid material appears to be useful in embolizing cerebral aneurysms safely.

Among five kinds of rat adhesion models tested, the following model was selected. The epigastric vein 2.5 cm from the midline of the abdomen was cut by sharp scissors, and the lateral side of the cut epigastric vein was ligated using a 3-0 silk suture. This model could be easily prepared and gave a rate of adhesion formation of 90%, which was useful for screening antiadhesive materials. For the kinetic study of tissue adhesion in this model, an injured site was covered with a non-degradable poly(vinyl alcohol) (PVA) film. The incidence rate of adhesion was 18%, when the PVA film covered the injured site for 2 days. This suggests that an antiadhesive barrier should cover the injured site for at least 2 days. The antiadhesion efficacy of cross-linked gelatin films were evaluated using this adhesion model. The UV cross-linked gelatin film which was designed to exist for 2 days but to disappear at day 3 in the rat abdominal cavity showed the highest antiadhesion efficacy.

Staple-line reinforcement buttresses made of bovine pericardium (BP), expanded polytetrafluoroethylene (ePTFE), and so on have been shown to be effective in preventing air leaks after stapled lung volume reduction operations, and some of them have been clinically utilized. However, each buttress suffers at least one disadvantage such as risk of viral infection and chronic inflammation. A new buttress was made using a poly(L-lactic acid-co-epsilon-caprolactone) film (L/C film) and its effectiveness as a staple-line reinforcement was examined by performing lung volume reduction operation on a canine model. Soft tissue responses to the buttress were compared with those to the BP strip and the absorbable behavior was studied. The L/C film buttress was flexible and thin enough to easily cut. Death of dogs, infection, acute and prolonged air leaks, and any complications related to its use were not observed. The tissue responses to the film were more mild and favorable than those to BP. The L/C film was absorbed after the staple line was covered by a connective tissue. The results described above suggest that the buttress made of an L/C film is a promising staple-line reinforcement material.

Endovascular neurosurgery is now becoming available as one of strategies for the treatment of cerebro-spinal arterio-venous malformations and aneurysms. For this treatment, a microcatheter is advanced into or close to a lesion and then an embolic material is administered through it to obliterate the lesion. N-butyl-2-cyanoacrylate (NBCA) has preferentially been used as an embolic material in Europe and America. However, its exceptionally strong adhesive force sometimes causes adhesion between the tip of the microcatheter and the artery. In this study, a new non-adhesive cyanoacrylate, isostearyl-2-cyanoacrylate (ISCA), was developed. It carries a long hydrophobic side isostearyl group with lower reactivity and adhesion than other cyanoacrylates. Its polymerization rate is, however, too low to obliterate a vascular lesion with a rapid blood flow. To increase the polymerization rate. ISCA was mixed with NBCA. As a result, the adhesive force of the mixture became extremely low, compared with that of NBCA. The viscosity of the mixture was low enough to allow its' use as an embolic material. Tissue reactions against the mixture was milder than those against NBCA. Radio-angiography became possible by mixing further with Lipiodol. The evaluation of this new embolic material with a rabbit renal artery showed that the obliteration effect of the mixture of ISCA and NBCA was excellent to use as an embolic material for clinical applications.

The present study was carried out in an attempt to make a gelatin film strongly bioadhesive by introducing free dangling aldehyde groups. When gelatin films were treated with 0.5M of glutaraldehyde (GA) solution at 60 degrees C, free aldehyde groups (up to 150 micromol/g) were introduced in the film. The bonding strength of GA-crosslinked gelatin films (GA gelatin films) with biological tissue was assessed using porcine skins. It was found that bonding strength increased with increasing aldehyde content in the film. The GA gelatin films had bonding strength as high as 250 gf/cm2 whereas the native gelatin film (before GA treatment) showed bonding strength of 40 gf/cm2. When the aldehyde groups introduced in the gelatin films were quenched with glycine or reduced by NaBH4, the films no longer demonstrated such high bonding strength. These facts suggest that a Schiff base was formed between the free dangling aldehyde in the GA gelatin films and the amino groups of the natural tissue, which strongly contributed to a marked bioadhesion.

Characteristics of a newly developed liquid material for embolization of AVM and its clinical experiments are reported. This liquid is composed of dissolving poly (2-hydroxyethyl methacrylate-co-methyl methacrylate) in Iopamiron with adding a small amount of ethyl alcohol. Thirty-two cases of AVM and 5 cases of spinal AVM were embolized with this liquid with good results. This liquid material is of low viscosity, not adhesive and not cytotoxic, therefore, it can be easily injected through a microcatheter, and can safely and effectively occlude AVM.

We have developed a new biodegradable scaffold that does not require any cell seeding to create an in-situ tissue-engineering vasculature (iTEV). Animal experiments were conducted to test its characteristics and long-term efficacy. An 8-mm tubular biodegradable scaffold, consisting of polyglycolide knitted fibers and an L-lactide and ε-caprolactone copolymer sponge with outer glycolide and ε-caprolactone copolymer monofilament reinforcement, was implanted into the inferior vena cava (IVC) of 13 canines. All the animals remained alive without any major complications until euthanasia. The utility of the iTEV was evaluated from 1 to 24 months postoperatively. The elastic modulus of the iTEV determined by an intravascular ultrasound imaging system was about 90% of the native IVC after 1 month. Angiography of the iTEV after 2 years showed a well-formed vasculature without marked stenosis or thrombosis with a mean pressure gradient of 0.51 ± 0.19 mmHg. The length of the iTEV at 2 years had increased by 0.48 ± 0.15 cm compared with the length of the original scaffold (2-3 cm). Histological examinations revealed a well-formed vessel-like vasculature without calcification. Biochemical analyses showed no significant differences in the hydroxyproline, elastin, and calcium contents compared with the native IVC. We concluded that the findings shown above provide direct evidence that the new scaffold can be useful for cell-free tissue-engineering of vasculature. The long-term results revealed that the iTEV was of good quality and had adapted its shape to the needs of the living body. Therefore, this scaffold would be applicable for pediatric cardiovascular surgery involving biocompatible materials.

This study evaluated the endothelial function and mechanical properties of tissue-engineered vascular autografts (TEVAs) constructed with autologous mononuclear bone marrow cells (MN-BMCs) and a biodegradable scaffold using a canine inferior vena cava (IVC) model. MN-BMCs were obtained from a dog and seeded onto a biodegradable tubular scaffold consisting of polyglycolide fiber and poly(L-lactide-co-epsilon-caprolactone) sponge. This scaffold was implanted in the IVC of the same dog on the day of surgery. TEVAs were analyzed biochemically, biomechanically, and histologically after implantation. When TEVAs were explanted and stimulated with acetylcholine at 1 month, they produced nitrates and nitrites dose dependently. N(G)-nitro-L-arginine methylester significantly inhibited these reactions. With stimulation by acetylcholine, factor VIII-positive cells of TEVAs produced endothelial nitric oxide synthase proteins, and the ratio of endothelial nitric oxide synthase/s17 mRNA was similar among native IVC and TEVAs 1 and 3 months after implantation. TEVAs had biochemical properties and wall thickness similar to those of native IVC at 6 months after implantation, and tolerated venous pressure well without any problems such as calcification. The number of inflammatory cells in TEVAs and the ratio of CD4/s17 mRNA decreased significantly with time. These results indicate that TEVAs are a biocompatible material with functional endothelial cells and biomechanical properties and do not have unwanted side effects.